Unveiling the potential of Pseudococcomyxa simplex: a stepwise extraction for cosmetic applications.

Microalgae are gaining attention as they are considered green fabrics able to synthesize many bioactive metabolites, with unique biological activities. However, their use at an industrial scale is still a challenge because of the high costs related to upstream and downstream processes. Here, a biorefinery approach was proposed, starting from the biomass of the green microalga Pseudococcomyxa simplex for the extraction of two classes of molecules with a potential use in the cosmetic industry. Carotenoids were extracted first by an ultrasound-assisted extraction, and then, from the residual biomass, lipids were obtained by a conventional extraction. The chemical characterization of the ethanol extract indicated lutein, a biosynthetic derivative of α-carotene, as the most abundant carotenoid. The extract was found to be fully biocompatible on a cell-based model, active as antioxidant and with an in vitro anti-aging property. In particular, the lutein-enriched fraction was able to activate Nrf2 pathway, which plays a key role also in aging process. Finally, lipids were isolated from the residual biomass and the isolated fatty acids fraction was composed by palmitic and stearic acids. These molecules, fully biocompatible, can find application as emulsifiers and softener agents in cosmetic formulations. Thus, an untapped microalgal species can represent a sustainable source for cosmeceutical formulations. KEY POINTS: • Pseudococcomyxa simplex has been explored in a cascade approach. • Lutein is the main extracted carotenoid and has antioxidant and anti-aging activity. • Fatty acids are mainly composed of palmitic and stearic acids.

Meiotic maturation failure in primary ovarian insufficiency: insights from a bovine model.

PURPOSE: Oocytes from women presenting primary ovarian insufficiency (POI) generate viable embryos at a lower rate than non-POI women, but the mechanisms responsible for the lower oocyte quality remain elusive. Due to the scarcity of human oocytes for research, animal models provide a promising way forward. We aimed at investigating the molecular events characterizing final maturation in POI oocytes in a well-defined POI-like bovine model. METHODS: Single-cell RNA-sequencing of bovine control and POI-like, GV, and MII oocytes (n = 5 per group) was performed. DEseq2 was used to identify differentially expressed genes. Further, a Gene set enrichment analysis and a transcriptomic meta-analysis between bovine and human oocytes were performed. RESULTS: In control cows, we found 2223 differentially expressed genes between the GV and MII stages. Specifically, the affected genes were related to RNA processing and transport, protein synthesis, organelle remodeling and reorganization, and metabolism. The meta-analysis with a set of young human oocytes at different maturation stages revealed 315 conserved genes through the GV-MII transition in cows and humans, mostly related to meiotic progression and cell cycle. Gene expression analysis between GV and MII of POI-like oocytes showed no differences in terms of differentially expressed genes, pointing towards a substantial failure to properly remodel the transcriptome in the POI model, and with the clustering analysis indicating that the cow's genetic background had a higher impact than the oocyte's maturation stage. CONCLUSION: Overall, we have identified and characterized a valuable animal model of POI, paving the way to identifying new molecular mechanisms involved in POI.

Tetraselmis chuii Edible Microalga as a New Source of Neuroprotective Compounds Obtained Using Fast Biosolvent Extraction.

Tetraselmis chuii is an EFSA-approved novel food and dietary supplement with increasing use in nutraceutical production worldwide. This study investigated the neuroprotective potential of bioactive compounds extracted from T. chuii using green biobased solvents (ethyl acetate, AcOEt, and cyclopentyl methyl ether, CPME) under pressurized liquid extraction (PLE) conditions and supercritical fluid extraction (SFE). Response surface optimization was used to study the effect of temperature and solvent composition on the neuroprotective properties of the PLE extracts, including anticholinergic activity, reactive oxygen/nitrogen species (ROS/RNS) scavenging capacity, and anti-inflammatory activity. Optimized extraction conditions of 40 °C and 34.9% AcOEt in CPME resulted in extracts with high anticholinergic and ROS/RNS scavenging capacity, while operation at 180 °C and 54.1% AcOEt in CPME yielded extracts with potent anti-inflammatory properties using only 20 min. Chemical characterization revealed the presence of carotenoids (neoxanthin, violaxanthin, zeaxanthin, α- and ß-carotene) known for their anti-cholinesterase, antioxidant, and anti-inflammatory potential. The extracts also exhibited high levels of omega-3 polyunsaturated fatty acids (PUFAs) with a favorable ω-3/ω-6 ratio (>7), contributing to their neuroprotective and anti-inflammatory effects. Furthermore, the extracts were found to be safe to use, as cytotoxicity assays showed no observed toxicity in HK-2 and THP-1 cell lines at or below a concentration of 40 µg mL-1. These results highlight the neuroprotective potential of Tetraselmis chuii extracts, making them valuable in the field of nutraceutical production and emphasize the interest of studying new green solvents as alternatives to conventional toxic solvents.

Specific processing of meiosis-related transcript is linked to final maturation in human oocytes.

Human meiosis in oocytes entails an intricate regulation of the transcriptome to support late oocyte growth and early embryo development, both crucial to reproductive success. Currently, little is known about the co- and post-transcriptional mRNA processing mechanisms regulating the last meiotic phases, which contribute to transcriptome complexity and influence translation rates. We analyzed gene expression changes, splicing and pre-mRNA processing in an RNA sequencing set of 40 human oocytes at different meiotic maturation stages, matured both in vivo and in vitro. We found abundant untranslated region (UTR) processing, mostly at the 3' end, of meiosis-related genes between the germinal vesicle (GV) and metaphase II (MII) stages, supported by the differential expression of spliceosome and pre-mRNA processing related genes. Importantly, we found very few differences among GV oocytes across several durations of IVM, as long as they did not reach MII, suggesting an association of RNA processing and successful meiosis transit. Changes in protein isoforms are minor, although specific and consistent for genes involved in chromosome organization and spindle assembly. In conclusion, we reveal a dynamic transcript remodeling during human female meiosis, and show how pre-mRNA processing, specifically 3'UTR shortening, drives a selective translational regulation of transcripts necessary to reach final meiotic maturation.

How do the activity patterns of people with chronic pain influence the empathic response of future health professionals: an experimental study.

Empathy in healthcare professionals is associated with better treatment outcomes and higher satisfaction among patients with chronic pain. Activity patterns play an essential role in the adjustment of these patients and, as a pain behaviour, may have a communicative function and elicit distinct empathic responses. This study investigated whether the activity pattern profiles characteristic of these patients had differential effects on the empathic response (empathic distress and compassion/sympathy) of future healthcare professionals. Healthcare professionals should improve their knowledge about the role of different activity patterns in the well-being of people with chronic pain and receive specific training in empathic skills. We controlled for several variables that could affect the empathic response (sex, age, academic degree, previous experience of chronic pain, and dispositional empathy).A total of 228 undergraduates performed an experimental task using vignettes depicting four activity pattern profiles displayed by people with chronic pain and completed questionnaires measuring dispositional and situational empathy. We conducted a MANCOVA analysis.Undergraduates showed more compassion/sympathy toward the medium cycler profile than toward the doer profile. Participants' age was associated with empathic distress. Sex, academic degree, and previous experiences with chronic pain were not associated with their empathic response to the vignettes. Dispositional perspective-taking and empathic concern were significantly associated with compassion/sympathy responses, and personal distress was significantly associated with empathic distress.Activity pattern profiles may have a communicative function and elicit different empathic responses toward people with chronic pain. Individual differences in dispositional empathy play an important role on situational empathic responses.

An Alternative Exploitation of Synechocystis sp. PCC6803: A Cascade Approach for the Recovery of High Added-Value Products.

Microalgal biomass represents a very interesting biological feedstock to be converted into several high-value products in a biorefinery approach. In this study, the cyanobacterium Synechocystis sp. PCC6803 was used to obtain different classes of molecules: proteins, carotenoids and lipids by using a cascade approach. In particular, the protein extract showed a selective cytotoxicity towards cancer cells, whereas carotenoids were found to be active as antioxidants both in vitro and on a cell-based model. Finally, for the first time, lipids were recovered from Synechocystis biomass as the last class of molecules and were successfully used as an alternative substrate for the production of polyhydroxyalkanoate (PHA) by the native PHA producer Pseudomonas resinovorans. Taken together, our results lead to a significant increase in the valorization of Synechocystis sp. PCC6803 biomass, thus allowing a possible offsetting of the process costs.

ANF therapy® for pain management, feasibility, satisfaction, perceived symptom reduction and side effects: a real-world multisite observational study.

[Purpose] Non-invasive and drug-free interventions for pain are being developed. One of them is ANF (which stands for "Amino Neuro Frequency") Therapy®, which consists in the application of carbonized metal devices on a patient's skin. We aimed to: 1) test perceived changes in pain intensity after ANF application, 2) record frequency and severity of side effects, 3) assess clinician and patient satisfaction, 4) explore effects on swelling and range of motion (ROM). [Participants and Methods] In this real-world multisite observational study, N=113 physical therapists in 45 countries, applied ANF to N=1,054 patients (Mage=45.2, 56.2% female) with pain complaints. Demographic data, pain intensity (NRS-11), effects of ANF on swelling and ROM, clinician and patient satisfaction and side effects were collected. [Results] Main pain locations were: low back (14.9%), knee (12.4%), neck (10%), and shoulder (9.6%). Pre-treatment pain intensity was high (Mean=7.6, SD=1.9). It significantly decreased post-treatment (Mean=3.1, SD=2.0), t(1053)=7.25, with a large effect size (Cohen's d=2.2). Swelling decreased and ROM increased. Average satisfaction with ANF was 92/100. Patients often experienced mild side effects (42.3%): dry mouth, headache and fatigue. [Conclusion] Results show large effect sizes, high satisfaction, and mild and short-term side effects. This is very promising but should be interpreted with caution considering the study limitations.

Study of the potential neuroprotective effect of Dunaliella salina extract in SH-SY5Y cell model.

Alzheimer's disease (AD) is the most common form of dementia caused by a progressive loss of neurons from different regions of the brain. This multifactorial pathophysiology has been widely characterized by neuroinflammation, extensive oxidative damage, synaptic loss, and neuronal cell death. In this sense, the design of multi-target strategies to prevent or delay its progression is a challenging goal. In the present work, different in vitro assays including antioxidant, anti-inflammatory, and anti-cholinergic activities of a carotenoid-enriched extract from Dunaliella salina microalgae obtained by supercritical fluid extraction are studied. Moreover, its potential neuroprotective effect in the human neuron-like SH-SY5Y cell model against remarkable hallmarks of AD was also evaluated. In parallel, a comprehensive metabolomics study based on the use of charged-surface hybrid chromatography (CSH) and hydrophilic interaction liquid chromatography (HILIC) coupled to high-resolution tandem mass spectrometry (Q-TOF MS/MS) was applied to evaluate the effects of the extract on the metabolism of the treated cells. The use of advanced bioinformatics and statistical tools allowed the identification of more than 314 metabolites in SH-SY5Y cells, of which a great number of phosphatidylcholines, triacylglycerols, and fatty acids were significantly increased, while several phosphatidylglycerols were decreased, compared to controls. These lipidomic changes in cells along with the possible role exerted by carotenoids and other minor compounds on the cell membrane might explain the observed neuroprotective effect of the D. salina extract. However, future experiments using in vivo models to corroborate this hypothesis must be carried out.

Intolerance of Uncertainty Moderates the Relationship between Catastrophizing, Anxiety, and Perceived Pain in People with Chronic Nononcological Pain.

OBJECTIVE: Substantial empirical evidence has shown that intolerance of uncertainty is a central transdiagnostic feature in psychopathology and it has been suggested to be a pain-related psychological factor contributing to the experience of chronic pain. However, research in this area is virtually nonexistent. The objective of this study was to investigate associations between pain severity, catastrophizing, and anxiety in people with chronic nononcological pain, while assuming that intolerance of uncertainty moderates these relationships. METHODS: A convenience sample of 188 individuals with nononcological chronic pain (157 women and 32 men) participated in the study. We investigated the moderated mediation of intolerance of uncertainty between anxiety and catastrophizing and between catastrophizing and pain intensity. RESULTS: The full moderated mediation model accounted for significant variance in pain intensity (R2 = 0.148, P< .001). Intolerance of uncertainty significantly moderated the interaction between anxiety and catastrophizing (B = 0.039, SE = 0.012, 95% CI [0.015, 0.063]) and between catastrophizing and pain intensity (B = -0.034, SE = 0.010, 95% CI [-0.054, -0.014]). Anxiety and intolerance of uncertainty did not interact in predicting catastrophizing, although an interaction effect was found between intolerance of uncertainty and catastrophizing in predicting pain intensity. CONCLUSION: This study is the first to address the interrelationship of intolerance of uncertainty, catastrophizing, and anxiety in relation to perceived pain intensity. The current findings support intolerance of uncertainty as a relevant psychological variable that is distinct from other relevant constructs in the setting of pain research and treatment.

Efficient generation of embryonic stem cells from single blastomeres of cryopreserved mouse embryos in the presence of signalling modulators.

CONTEXT: Derivation of embryonic stem cells (ESC) from single blastomeres is an interesting alternative to the use of whole blastocysts, but derivation rates are lower and the requirements for successful ESC obtention are still poorly defined. AIMS: To investigate the effects of embryo cryopreservation and of signalling modulators present during embryo culture and/or ESC establishment on ESC derivation efficiency from single 8-cell mouse blastomeres. METHOD: Fresh and cryopreserved 2-cell embryos were cultured and biopsied at the 8-cell stage. Single blastomeres were cultured in the presence of 2i or R2i cocktails, with or without adrenocorticotropic hormone (ACTH). We analysed ESC derivation efficiencies and characterised pluripotency genes expression and karyotype integrity of the resulting lines. We also evaluated the impact of embryo preculture with R2i on epiblast cell numbers and derivation rates. KEY RESULTS: The ESC generation was not compromised by embryo cryopreservation and ACTH was dispensable under most of the conditions tested. While 2i and R2i were similarly effective for ESC derivation, R2i provided higher karyotype integrity. Embryo preculture with R2i yielded increased numbers of epiblast cells but did not lead to increased ESC generation. CONCLUSIONS: Our findings help to define a simplified and efficient procedure for the establishment of mouse ESC from single 8-cell blastomeres. IMPLICATIONS: This study will contribute to improving the potential of this experimental procedure, providing a tool to investigate the developmental potential of blastomeres isolated from different embryonic stages and to reduce the number of embryos needed for ESC derivation.

Emerging Lipids from Arecaceae Palm Fruits in Brazil.

Arecaceae palm tree fruits (APTFs) with pulp or kernel rich in oil are widely distributed in six Brazilian biomes. APTFs represent a great potential for the sustainable exploitation of products with high added value, but few literature studies have reported their properties and industrial applications. The lack of information leads to underutilization, low consumption, commercialization, and processing of these fruit species. This review presents and discusses the occurrence of 13 APTFs and the composition, physicochemical properties, bioactive compounds, and potential applications of their 25 oils and fats. The reported studies showed that the species present different lipid profiles. Multivariate analysis based on principal component analysis (PCA) and hierarchical cluster analysis (HCA) indicated a correlation between the composition of pulp and kernel oils. Myristic, caprylic, capric, and lauric acids are the main saturated fatty acids, while oleic acid is the main unsaturated. Carotenoids and phenolic compounds are the main bioactive compounds in APTFs, contributing to their high oxidative stability. The APTFs oils have a potential for use as foods and ingredients in the cosmetic, pharmaceutical, and biofuel industries. However, more studies are still necessary to better understand and exploit these species.

Electroless Palladium-Coated Polymer Scaffolds for Electrical Stimulation of Osteoblast-Like Saos-2 Cells.

Three-dimensional porous scaffolds offer some advantages over conventional treatments for bone tissue engineering. Amongst all non-bioresorbable scaffolds, biocompatible metallic scaffolds are preferred over ceramic and polymeric scaffolds, as they can be used as electrodes with different electric field intensities (or voltages) for electric stimulation (ES). In the present work we have used a palladium-coated polymeric scaffold, generated by electroless deposition, as a bipolar electrode to electrically stimulate human osteoblast-like Saos-2 cells. Cells grown on palladium-coated polyurethane foams under ES presented higher proliferation than cells grown on foams without ES for up to 14 days. In addition, cells grown in both conditions were well adhered, with a flat appearance and a typical actin cytoskeleton distribution. However, after 28 days in culture, cells without ES were filling the entire structure, while cells under ES appeared rounded and not well adhered, a sign of cell death onset. Regarding osteoblast differentiation, ES seems to enhance the expression of early expressed genes. The results suggest that palladium-coated polyurethane foams may be good candidates for osteoblast scaffolds and demonstrate that ES enhances osteoblast proliferation up to 14 days and upregulate expression genes related to extracellular matrix formation.

Biocompatibility and Electrical Stimulation of Skeletal and Smooth Muscle Cells Cultured on Piezoelectric Nanogenerators.

Nanogenerators are interesting for biomedical applications, with a great potential for electrical stimulation of excitable cells. Piezoelectric ZnO nanosheets present unique properties for tissue engineering. In this study, nanogenerator arrays based on ZnO nanosheets are fabricated on transparent coverslips to analyse the biocompatibility and the electromechanical interaction with two types of muscle cells, smooth and skeletal. Both cell types adhere, proliferate and differentiate on the ZnO nanogenerators. Interestingly, the amount of Zn ions released over time from the nanogenerators does not interfere with cell viability and does not trigger the associated inflammatory response, which is not triggered by the nanogenerators themselves either. The local electric field generated by the electromechanical nanogenerator-cell interaction stimulates smooth muscle cells by increasing cytosolic calcium ions, whereas no stimulation effect is observed on skeletal muscle cells. The random orientation of the ZnO nanogenerators, avoiding an overall action potential aligned along the muscle fibre, is hypothesised to be the cause of the cell-type dependent response. This demonstrates the need of optimizing the nanogenerator morphology, orientation and distribution according to the potential biomedical use. Thus, this study demonstrates the cell-scale stimulation triggered by biocompatible piezoelectric nanogenerators without using an external source on smooth muscle cells, although it remarks the cell type-dependent response.

Selective Extraction of Piceatannol from Passiflora edulis by-Products: Application of HSPs Strategy and Inhibition of Neurodegenerative Enzymes.

Passiflora edulis by-products (PFBP) are a rich source of polyphenols, of which piceatannol has gained special attention recently. However, there are few studies involving environmentally safe methods for obtaining extracts rich in piceatannol. This work aimed to concentrate piceatannol from defatted PFBP (d-PFBP) by means of pressurized liquid extraction (PLE) and conventional extraction, using the bio-based solvents selected with the Hansen solubility parameters approach. The relative energy distance (Ra) between solvent and solute was: Benzyl Alcohol (BnOH) < Ethyl Acetate (EtOAc) < Ethanol (EtOH) < EtOH:H2O. Nonetheless, EtOH presented the best selectivity for piceatannol. Multi-cycle PLE at 110 °C was able to concentrate piceatannol 2.4 times more than conventional extraction. PLE exhibited a dependence on kinetic parameters and temperature, which could be associated with hydrogen bonding forces and the dielectric constant of the solvents. The acetylcholinesterase (AChE) and lipoxygenase (LOX) IC50 were 29.420 µg/mL and 27.682 µg/mL, respectively. The results reinforce the demand for processes to concentrate natural extracts from food by-products.

The relevance of psychological strength for physical and psychological well-being in trauma-exposed women.

Traumatic experiences have consistently been linked to poor health and well-being, particularly in women. Psychological factors have been theorized to directly affect the reporting of physical symptoms and perceptions of general health. Posttraumatic stress disorder (PTSD) has been proposed as a major pathway through which trauma affects health and emotion dysregulation. Trauma is considered to be a key psychological variable in the pathogenesis of PTSD. Fortunately, not all women who have experienced trauma manifest adverse effects. Resilience acts as a psychological protective variable following trauma. The present study tested a hypothetical model of the contribution of resilience, emotional dysregulation, and PTSD symptoms to physical and psychological well-being in a large sample of trauma-exposed women. A transversal study with 753 female participants is used. Structural modeling was used to test linear associations between variables. After experiencing trauma, resilience was negatively and significantly associated with emotional dysregulation, which, in turn, was positively associated with PTSD symptoms. Both resilience and PTSD symptoms were associated with physical and psychological well-being. The results suggest that resilience and emotional dysregulation are relevant to the health and well-being of women with PTSD symptoms and may help guide the development of psychological treatment in this group. Therefore, these findings may be relevant in promoting health and well-being in such women, and may help to identify individuals who would receive the most benefit from interventions addressing emotional regulation and psychological resilience.

The Relationship Between Experiential Avoidance and Posttraumatic Stress Symptoms: A Moderated Mediation Model Involving Dissociation, Guilt, and Gender.

Experiential avoidance, dissociation, and guilt have been shown to be associated with trauma exposure and to play an important role in explaining the development and maintenance of posttraumatic stress symptoms. However, there is a lack of studies that simultaneously address the relationship between these variables, which has never been studied within the framework of emotional processing theory. Furthermore, gender differences in traumatic victimization, posttraumatic stress symptoms, experiential avoidance, dissociation, and guilt have also been reported. Therefore, this study had a double aim: a) to assess the mediating roles of dissociation and guilt in the relationship between experiential avoidance and posttraumatic stress symptoms; and b) to investigate whether gender moderates any such relationship. The final sample comprised 683 undergraduate students (150 men and 533 women) with a history of exposure to traumatic events and with posttraumatic stress symptoms. Mediation and moderated mediation analyses were conducted. Dissociation and guilt independently mediated the association between experiential avoidance and posttraumatic stress symptoms: however, this mediation effect was not moderated by gender. The findings suggest that interventions aimed at controlling psychological variables linked to PTSD (i.e. experiential avoidance, dissociation, and guilt) may be of help to both men and women.

Chemical characterization of leaves and calli extracts of Rosmarinus officinalis by UHPLC-MS.

Rosmarinus officinalis L. (Lamiaceae) is an aromatic plant widely popular mainly due to its uses in traditional medicine as an anti-inflammatory, diuretic and antimicrobial, as well as in the prevention and treatment of diseases. These biological activities are mainly related to the presence of phenolic and terpenic compounds. This work reports a chemical profile analysis of extracts from leaves and calli of rosemary obtained by both pressurized liquid extraction and maceration. Chemical profiles were determined on calli extracts of 3, 6, 9, and 15 days of culture; chemical characterization and quantification of compounds was carried out using ultrahigh performance liquid chromatography-mass spectrometry. A total of 53 metabolites were identified in callus and 47 compounds in leaf extracts, of which 25 correspond to phenolic compounds, mainly flavonoids and flavones, 13 terpenes that include phenolic terpenes and one diterpenolactone, two glycosides which correspond to 6-O-caffeoyl-ß-D-fructofuranosil-(2→1)-α-D-glucopyranoside and primulaverin, an aromatic compound identified as fenantrenone and a growth regulator 12-hydroxy jasmonic acid. These results showed that undifferentiated rosemary cells accumulate the same compounds identified mainly in highly specialized tissues such as leaves. The plant cell culture supply the possibility of developing biotechnological processes to obtain compounds of commercial interest.

Compressed fluids and phytochemical profiling tools to obtain and characterize antiviral and anti-inflammatory compounds from natural sources.

Many natural compounds, found mainly in plants, are associated with the treatment of various diseases. The search for natural therapeutic agents includes compounds with antiviral and anti-inflammatory activities. Among the many steps involved in bioprospection, extraction is the first and most critical step for obtaining bioactive compounds. One of the main advantages of using compressed fluids extraction is the high quality of the final product obtained due to the use of green solvents, while the selectivity towards target compounds can be tuned by adjusting the process parameters, especially pressure, temperature and solvent characteristics. In this review, a discussion is provided on the power of compressed fluids, such as supercritical fluid extraction (SFE), pressurized liquid extraction (PLE) and subcritical water extraction (SWE) to obtain antiviral and anti-inflammatory compounds from natural sources. In addition, an adequate knowledge about the identity and quantity of the compounds present in the extract is essential to correlate biological activity with chemical composition. Phytochemical profiling tools used for identification and quantification of these bioactive natural compound are also discussed. It can be anticipated that after the current SARS-COV-2 pandemic, the search of new natural compounds with antiviral and anti-inflammatory activity will be a hot research topic, so, this review provides an overview on the technologies currently used that could help this research.

Application of compressed fluid-based extraction and purification procedures to obtain astaxanthin-enriched extracts from Haematococcus pluvialis and characterization by comprehensive two-dimensional liquid chromatography coupled to mass spectrometry.

The green microalga Haematococcus pluvialis has been widely studied due to its capacity to accumulate great amounts of astaxanthin, a high-value carotenoid with biological activities. In the present work, two green compressed fluid-based processes, pressurized liquid extraction (PLE) and supercritical antisolvent fractionation (SAF), are integrated to obtain an astaxanthin-enriched extract from this microalga. PLE was carried out using pressurized ethanol as solvent, for 20 min, at 10 MPa, and 50 °C as extraction temperature. Subsequently, the obtained extract was processed by SAF to further purify the carotenoid fraction. The SAF process was optimized using a 3-level factorial experimental design and considering three experimental variables: (i) CO2 pressure (10-30 MPa), (ii) percentage of water in the PLE extract (20-50%), and (iii) PLE extract/supercritical-CO2 flow rate ratio (0.0125-0.05). Total carotenoid content was evaluated in both extracts and raffinates. Best results were obtained at 30 MPa, 0.05 feed/SC-CO2 mass flow rate, and 20% (v/v) of water in the feed solution, achieving values of 120.3 mg g-1 carotenoids in extract (in the SAF extract fraction), which were significantly higher than those obtained in the original PLE extract. In parallel, a new fast two-dimensional comprehensive liquid chromatography (LC×LC) method was optimized to get the full carotenoid profile of these extracts in less than 25 min. This is the first time that the use of a C30 column is reported in an on-line LC×LC system. Graphical abstract.

Using vignettes to assess the contribution of goal adjustment in the adaptation to chronic musculoskeletal pain.

PURPOSE: The quality of life of individuals with chronic musculoskeletal pain partly depends on their capacity to adjust their personal goals. Vignettes have been rarely used to assess this ability. Therefore, this study aimed to test the relationships between vignettes assessing different goal strategies and chronic pain adaptation (i.e., daily functioning, pain-related impairment, and psychological well-being). METHODS: The sample comprised 258 individuals with chronic musculoskeletal pain who completed a series of questionnaires and vignettes. The vignettes presented a short description of a situation in which a person with chronic pain experienced a threat to a valued domain-specific goal and had to choose a possible goal management solution (i.e., goal persistence, flexibility reengagement, and disengagement). Hierarchical regression analyses were used to predict chronic pain adaptation using the selected vignette strategies as predictors. RESULTS: After controlling for age, sex, pain intensity, and the responses to the dispositional goal management scales, persistence, reengagement, and disengagement goal strategies presented in the case scenarios predicted daily functioning (p < .001). Persistence, flexibility, disengagement (p < .001), and reengagement (p < .05) predicted pain-related impairment. Persistence, disengagement (p < .001), and flexibility (p < .05) predicted psychological well-being scores. CONCLUSION: The use of vignettes could be useful to assess goal adjustment because this methodology enables respondents to provide more context-specific responses. The results of this approach could be used to improve clinical practice aimed at helping people with chronic musculoskeletal pain to better cope with this health condition.